Stock Number: 006390 Strain Name: B6;129-Sncatm1Sud Sncbtm1.1Sud/J

Allele: Sncbtm1.1Sud Protocol Name: Sncbtm1.1Sud Method: Standard PCR Version: 1.1 Created: 26-June -2008 Updated: 24-September-2009

Expected Results: Mutant = 420 bp Heterozygote = 420 bp and 500 bp Wild type=500 bp

Gel Image

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Gel Information: Separated by gel electrophoresis on a 1.5% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR6429 - CAG CGT CTG GCA GCC GCT GAT TGG TGG - Wild type oIMR6430 - GTT GGT TGG AGG TGC GGT GCG TGA GG - Wild type oIMR6431 - CGT CTG GCA GCC GCT GAT TGG TGG - Mutant oIMR8574 - GAG CGC GCG CGG CGG AGT TGT TGA C - Mutant

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 2.76 - 2.76 10 X PCR Buffer II (ABI) 1.20 1.00 X 1.20 25 mM MgCl2 0.96 2.00 mM 0.96 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR6429 0.60 1.00 uM 0.60 20 uM oIMR6430 0.60 1.00 uM 0.60 20 uM oIMR6431 0.60 1.00 uM 0.60 20 uM oIMR8574 0.60 1.00 uM 0.60 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 5 U/ul Taq DNA Polymerase 0.06 0.03 U/ul 0.06 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 70 1 min - 4 72 1 min repeat steps 2-4 for 35 cycles 5 72 2 min - 6 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 006390 B6;129-Sncatm1Sud Sncbtm1.1Sud/J 008132 STOCK Tg(THY1-Snca)M1mSud/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.